Key Laboratory of Applied Surface and Colloid Chemistry, Ministry of Education, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi'an 710062, China.
Biosens Bioelectron. 2013 May 15;43:231-6. doi: 10.1016/j.bios.2012.12.004. Epub 2012 Dec 22.
A label-free fluorescent AND logic gate has been developed utilizing ion-tuned configuration conversion of DNA probe with K(+) and Pb(2+) as two inputs. A well-designed hairpin DNA which is composed of a poly-G loop and a GR-5 DNAzyme stem serves as a recognition probe, and an derivative of aloe-emodin (AED) was designed and synthesized as signal probe. In the presence of Pb(2+), the substrate strand of DNAzyme is irreversibly and specifically cleaved at the cleavage site, which made the poly-G loop form G-quadruplex in the presence of a constant concentration of K(+). Such a structural change significantly affects the spectral behaviors of AED, which can be explored to ultra-sensitively detect Pb(2+) with a limit of detection of 22.8pM. By combing the high specificity of hairpin DNA and GR-5 DNAzyme, Pb(2+) can be highly selectively detected even when coexisted with other metal ions. Circular dichroism (CD), UV-vis absorption spectrometry and fluorescence polarization (FP) measurements further verified the reliability and reasonability of the sensing mechanism. Therefore, it provides a simple and label-free approach to detect ions with high sensitivity and specificity, and promises to provide a solid sensing platform for the detection of targets by altering the specific sequence of nucleic acid probe.
一种无标记荧光 AND 逻辑门已被开发出来,利用 K(+)和 Pb(2+)作为两个输入,通过离子调谐 DNA 探针的构象转换来实现。一个精心设计的发夹 DNA 由一个多 G 环和一个 GR-5 DNA 酶茎组成,用作识别探针,并且设计并合成了大黄素衍生物(AED)作为信号探针。在 Pb(2+)存在下,DNA 酶的底物链在切割位点不可逆且特异性地被切割,这使得在恒定浓度的 K(+)存在下多 G 环形成 G-四链体。这种结构变化显著影响 AED 的光谱行为,可用于超灵敏地检测 Pb(2+),检测限低至 22.8pM。通过结合发夹 DNA 和 GR-5 DNA 酶的高特异性,即使存在其他金属离子,也可以高度选择性地检测 Pb(2+)。圆二色性(CD)、紫外可见吸收光谱和荧光偏振(FP)测量进一步验证了传感机制的可靠性和合理性。因此,它提供了一种简单且无标记的方法,用于以高灵敏度和特异性检测离子,并且有望通过改变核酸探针的特定序列来提供用于检测靶标的稳健传感平台。